Objective:To investigate the effect of neonatal dog peripheral mononuclear cells on left ventricular function, haemodynamic, and infarcted size when transplanted into infarcted myocardium. Methods: Mononuclear cells from neonatal dogs were isolated and purified. The descending coronary arteries of adult dogs were ligated and the cells were injected into the border zone of the infarcted region. The control group was injected with non serum culture medium. Left ventricular pressure, d p /d t max , and power of LV were measured during and 30 d after operation. Echocardiogram(HP Sonos 5500, 2.5 MHz) were performed promptly and 30 d after operation to determine the changes of left ventricular function. Infarcted size, capillary density and cardiac muscle width were measured 30 d after operation. Results: LV pressure decreased in both groups, there were no marked differences between the 2 groups. LV pressure and d p /d t max of transplant group were greater than those of control group 30 d after operation. The systolic and diastolic functions were preserved after mononuclear cell transplantation. Infarcted sizes were lower and capillary density was greater in transplant group than those of control group. Conclusion: Transplantation of neonatal dog peripheral mononuclear cells into the infarcted regions can increase the capillary density and decrease infarcted size, preserving left ventricular function.

Objective The olecranon is likely to become shorter following the treatment of its comminuted fractures, which will lead to the compromise of the function of elbow joint. So through the cadaver experiment, introduce the method of making different thickness and direction in osteotomy so as to intimate the shape change of the olecranon after the comminuted fractures, and explore its effect on the flexion-extension function of the elbow joint. Methods Through three cadaver(six arms), osteotomy at 25 mm below the olecranon process was made horizontally and fixed with two screws temporarily, furthermore, the bone fragment of 1 mm, 3 mm and 5 mm in length was resected in order, then specimens were divided into two groups: In the first group, osteotomy was made up to 7 mm and 8 mm continually, and the changes of the range of the elbow joint movement were measured; in the second group, the additional wedge osteotomy of 5 mm and 7 mm was performed respectively, then the outcomes between the two groups were compared. Results If the osteotomized bone was within 3 mm, shortening internal fixation was satisfactory for the reconstruction of the elbow joint function. However, in cases of osteotomized bone of 5 mm, the extension function would be limited as the loss of the trochlear notch is too much. In order to keep the normal range motion of the elbow, the dorsal cortex distal to osteotomy should be scarified about 3 mm for the wedge osteotomy. When the shortage attained to 7 mm, the elbow instability would occur, even if advanced wedge osteotomy was accomplished. Conclusion In cases of the comminuted olecranon fractures, if the osteotomy is made within 3 mm, the shortening fixation is appropriate; if it has not exceeded 5 mm, the fracture should be treated with advanced wedge osteotomy tilting back to keep the radian of the trochlea; and if it has reached 7 mm, bone grafting is necessary for recovering of the flexion-extension of the elbow joint.

Objective To introduce a new bilateral triceps brachii approach for the treatment of intercondylar fractures of the humerus, and explore the possibility for the operation without injuring the mechanism of extension of the elbow. Methods With fresh cadaver specimens, the triceps brachii was stripped off from the distal end of the humerus, the muscle belly was elevated and retracted bilaterally, then the height was recorded, and the exposure of the distal humerus was observed, especially to the trochlear region when the elbow were flexed at 15? , 30? , 45? , 60? , 80? respectively. Results Through the cadaver specimen observation and the clinical application, the reduction and fixation of the intercondylar fractures of humerus should be performed when the elbow is flexed at 45?－ 60?， at 15?－ 30? flexion, fracture over the supracondylar can be treated and finally at 80? flexion, the reduction of the trochlear region can be examined. Conclusion This bilateral approach through the triceps brachii is suitable for the treatment both of the intercondylar and epicondylar fractures.

Objective: To investigate the effects of glycogen synthase kinase-3β (GSK3β)/eukaryotic extension factor kinase 2 (eEF2K) signaling pathway on the process of pulmonary fibrosis through in vivo experiments, and find new ideas for clinical treatment of pulmonary fibrosis. Methods: The pulmonary fibrosis model of C57BL/6 male mice was induced by bleomycin with intratracheal injection at the dose of 2 mg/kg. After 14 days of modeling, animals were divided into model group, negative inhibition group and inhibition group (n=5 for each group), and control group was not processed. The inhibition group was treated with TDZD-8 (4 mg/kg) after modeling, the negative inhibition group was given DMSO solution after modeling, and the samples were collected after 28 days. Hematoxylin-eosin staining method was used to detect lung fibrosis in mice and scored according to Ashcroft scale. Expression levels of GSK3β, p-GSK3β, eEF2K, p-eEF2K (Ser70, Ser392, Ser470), precursor protein of matrix metalloproteinase-2 (pro-MMP-2), matrix metalloproteinase-2 (MMP-2), collagen I (Col I), collagen Ⅲ (Col Ⅲ) and α-smooth muscle actin (α-SMA) were detected by Western blot. Results: Compared with control group, the fibrosis score was up-regulated, the expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were increased, while that of eEF2K was decreased in model group (P＜0.05). Compared with model group, the fibrosis score, expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were decreased, but the expression level of eEF2K was increased in inhibition group (P＜0.05). Conclusion: GSK3β can activate eEF2K by phosphorylation at the sites of Ser70, Ser392 and Ser470, increase the contents of fibrosis indicators, promote the formation of pulmonary fibrosis, and aggravate lung tissue lesions.
Animals ; Collagen ; Collagen Type I ; Elongation Factor 2 Kinase/metabolism* ; Eukaryota/metabolism* ; Fibrosis ; Glycogen Synthase Kinase 3 beta ; Male ; Matrix Metalloproteinase 2/metabolism* ; Mice ; Mice, Inbred C57BL ; Pulmonary Fibrosis/chemically induced* ; Signal Transduction

To enhance in vitro dissolution of Cur by preparing Cur solid dispersions. The ability of HPMCAS-HF,HPMCAS-MF,HPMCAS-LF and PVPK30 to maintain supersaturated solution was investigated by supersaturation test. Amorphous solid dispersions were prepared by the solvent-evaporation method. The prepared samples were characterized using infrared spectroscopy( IR) and differential scanning calorimetry( DSC),and in vitro dissolution was investigated. DSC and IR results showed that in 1 ∶3 and 1 ∶9 solid dispersions,Cur was amorphously dispersed in the carrier,and the interaction existed between drug and carrier. The supersaturation test showed that the order of the ability of polymer to inhibit crystallization of Cur was MF>HF>LF>K30. The dissolution results showed that Cur-K30 amorphous solid dispersion had the highest drug release rate; Cur-K30 and Cur-LF amorphous solid dispersions had a quicker but not stable dissolution rate,and the drug concentration decrease after 4 h; Cur-MF and Cur-HF solid dispersions had a low dissolution,which however increased steadily,attributing to the strong ability of the polymers to inhibit the crystallization of Cur. HPMCAS could inhibit the degradation of Cur better than K30,especially MF and HF. The amorphous solid dispersions of cur significantly enhanced the dissolution of Cur and improved the chemical stability of Cur. This study can provide a basis for the rational selection of the polymer used for Cur solid dispersion.
Chemistry, Pharmaceutical ; Curcumin ; chemistry ; Drug Stability ; Methylcellulose ; analogs & derivatives ; chemistry ; Polymers ; Solubility

BACKGROUND:Injection of isoproterenol is known to induce proliferation and hypertrophy of acinar cells in rodent salivary glands. However, the clonal proliferation ability of stem/progenitor cells of salivary glands by isoproterenol remains unclear.
OBJECTIVE:To study the proliferation and activation ability of stem/progenitor cells of submandibular gland with colony assay by intraperitoneal injection of isoproterenol.
METHODS:Sprague-Dawley rats were randomly divided into two groups, isoproterenol and control groups, respectively intraperitonal y injected with isoproterenol and normal saline for 5 consecutive days. The gland tissues were harvested, and the stem/progenitor cells of submandibular gland were obtained by enzyme digestion in vitro. The number of clonal colonies of each group was analyzed. The larger colony cells were col ected for immunohistochemistry staining with CD90.1, laminin andα6β1.
RESULTS AND CONCLUSION:The number of middle and low proliferative potential colony-forming cells was less but high proliferative potential colony forming cells were significantly more in isoproterenol group compared with control group (P<0.05). However, there was no significant difference in the total number of the colonies between two groups (P>0.05). The high proliferative potential colony forming cells were positive for CD90.1, laminin andα6β1. Results showed that isoproterenol treatment model cannot increase the cellnumber, but enhance the proliferation ability of stem/progenitor cells from the submandibular gland.

Objective: To establish a method for the determination of chlorogenic acid, caffeic acid, and 6-O-p-coumaroyl-D-glucopyranose in Lygodii Herba. Methods: The determination was developed on Phenomenex C18 column (250 mm × 4.6 mm, 5 μm), methanol-acetonitrile-1% glacial acetic acid (5:5:90) was used as mobile phase, the detective wavelength was 325 nm, and the column temperature was 35 ℃. Results: The linear ranges of chlorogenic acid, caffeic acid, and 6-O-p-coumaroyl-D-glucopyranose were 0.368-2.760, 0.064-0.480, and 0.102-0.765 μg, respectively. The average recoveries were 101.48% (RSD = 1.44%), 99.56% (RSD = 0.92%), and 101.73% (RSD = 1.62%), respectively. Conclusion: The method is simple, accurate, and highly reproducible, which could be used for the quality control of Lygodii Herba.

Objective To establish method for simultaneous determination of hesperidin, cinnamaldehyde and eugenol in Chunyang Zhengqi capsules by high performance liquid chromatography. Methods The column was Agilent PorosheⅡ 120 EC-C₁₈ (4.6 mm×150 mm, 4 μm). The mobile phase was acetonitrile-water with gradient elution. The column temperature was 35℃. The flow rate was 1.0 ml/min, and the detection wavelength was 284 nm. Results The methodological verification showed that hesperidin, cinnamaldehyde and eugenol had a good linearity (r≥0.999 9). The precisions were less than 2.0%. The average recovery was between 98.0% and 101.9%. The stability and repeatability of RSD were also less than 3.0%, which met the requirements of method validation. Conclusion The method is simple, stable, reproducible and accurate, which could be used to the quality control of Chunyang Zhengqi capsules.

A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL^-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.

OBJECTIVE: To investigate the effects of quercetin on the apoptosis of platelets and to analyze the intrinsic mechanism. METHODS: Firstly, the effects of quecetin on the apoptosis of platelets was detected by flow cytometry. Secondly, Western blot was used to detect the expression of apoptosis-related proteins in the platelets treated with quercetin for 2 and 4 day. RESULTS: By flow cytometry, it was found that the apoptosis of platelets in the quercetin-treated group (2, 4 and 8 μmol/L) was inhibited, the apoptosis rate of platelets in 2, 4 and 8 μmol/L quercetin group was 3.12%±0.32%, 2.89%±0.15% and 2.31%±0.28%, respectively, which were signigicantly lover than that in control group (P＜0.01). With the increase of quecetin concentration, the proportion ratio of platelets significantly decreased in a concentration-dependent manner（r=-0.9985）. Similar results were observed on the 4th day. Western blot showed that the treatment with quercetin (2, 4 and 8 μmol/L) promoted the expression of anti-apoptotic protein BCL-2, inhibited the expression of pro-apoptotic protein BAX, resulting in a significant increase in the ratio of BCL-2/BAX (P＜0.01), thereby inhibiting the apoptosis of platelets. Similar results were observed on the 4th day. CONCLUSION Quercetin can inhibit platelet apoptosis by increasing the ratio of apoptosis-related protein BCL-2/BAX in a concentration-dependent manner.
Apoptosis ; Apoptosis Regulatory Proteins ; Blood Platelets ; Quercetin